Phillip A. Moore

Propofol versus thiopental: effects on peri‐induction intraocular pressures in normal dogs

OBJECTIVE To determine the effects of propofol or thiopental induction on intraocular pressures (IOP) in normal dogs. STUDY DESIGN Prospective randomized experimental study. ANIMALS Twenty-two random-source dogs weighing 19.5 +/- 5.3 kg. METHODS Dogs were randomly assigned to receive propofol 8 mg kg(-1) IV (group P) or thiopental 18 mg kg(-1) IV (group T) until loss of jaw tone. Direct arterial blood pressure, arterial blood gasses, and IOP were measured at baseline, after pre-oxygenation but before induction, before endotracheal intubation, and after intubation. RESULTS There were no significant differences between groups with regard to weight, body condition score, breed group, or baseline or before-induction IOP, arterial blood pressure, or blood gases. The baseline IOP was 12.9 mmHg. Before endotracheal intubation, IOP was significantly higher compared to baseline and before induction in dogs receiving propofol. After intubation with propofol, IOP was significantly higher compared to thiopental and was significantly higher compared to before induction. After intubation, IOP was significantly lower compared to before intubation in dogs receiving thiopental. Propofol increased IOP before intubation by 26% over the before-induction score and thiopental increased IOP by 6% at the same interval. The IOP in group P remained 24% over the before induction score whereas thiopental ultimately decreased IOP 9% below baseline after intubation. There was no significant relationship between any cardiovascular or blood gas parameter and IOP at any time. There was no significant relationship between the changes in any cardiovascular or blood gas parameter and the changes in IOP between time points. CONCLUSIONS AND CLINICAL RELEVANCE Propofol caused a significant increase in IOP compared to baseline and thiopental. Thiopental caused an insignificant increase in IOP which decreased after intubation. Propofol should be avoided when possible in induction of anesthesia in animals where a moderate increase in IOP could be harmful.

Characterization of cytokines associated with Th17 cells in the eyes of horses with recurrent uveitis

OBJECTIVE Equine recurrent uveitis (ERU) is a spontaneous disease that is the most common cause of blindness in horses, affecting up to 15% of the horse population. Th17 cells are a major cell population driving the pathogenesis in several mouse models of autoimmune inflammation, including experimental autoimmune uveitis. The purpose of this study is to investigate the role a Th17 cell-mediated response plays in the pathogenesis of ERU. PROCEDURE Banked, Davidsons-fixed equine globes histopathologically diagnosed with ERU (n = 7) were compared immunohistochemically with healthy control globes (n = 7). Immunohistochemical staining was performed using a pan-Leptospira antibody and antibodies against IL-6, IL-17, and IL-23. Additionally, immunostaining was performed for T-cell (CD3) and B-cell (CD79α) markers. Specificity of immunoreactivity was confirmed by western blot analysis. RESULTS Immunohistochemical staining was positive for IL-6, IL-17, and IL-23 within the cytoplasm of nonpigmented ciliary epithelial cells and mononuclear inflammatory cells infiltrating the iris, and ciliary body of ERU horses (n = 7) but negative in controls (n = 7). ERU-affected eyes were CD3 positive (n = 7) and CD79α negative (n = 7). Staining for Leptospira was negative in all ERU and control globes. CONCLUSIONS Strong immunoreactivity for IL-6, IL-17, and IL-23, in conjunction with the fact that T lymphocytes are the predominating inflammatory cells present in ERU, suggests that IL-17-secreting helper T-cells play a role in the pathogenesis of ERU. These findings suggest that horses with ERU may serve as a naturally occurring animal model for autoimmune uveitis.

Presence of chelonid fibropapilloma-associated herpesvirus in tumored and non-tumored green turtles, as detected by polymerase chain reaction, in endemic and non-endemic aggregations, Puerto Rico

Fibropapillomatosis (FP), a transmissible neoplastic disease of marine turtles characterized by a likely herpesviral primary etiology, has emerged as an important disease in green sea turtles (Chelonia mydas) over the past three decades. The objectives of this study were to determine the suitability of three different chelonid fibropapilloma-associated herpesvirus (CFPHV) gene targets in polymerase chain reaction (PCR) assays of affected tissues; to explore the presence of CFPHV in non-affected skin from turtles with and without tumors; and to better understand tissue localization of the CFPHV genome in a tumor-free turtle by evaluating CFPHV presence in microanatomic tissue sites. Two aggregations of green sea turtles (Chelonia mydas) in Puerto Rico were evaluated, with six sampling intervals over the three-year period 2004–2007. Primary and nested PCR for three different herpesviral gene targets- DNA polymerase, capsid maturation protease, and membrane glycoprotein B- were performed on 201 skin biopsies taken from 126 turtles with and without external tumors. Laser capture microdissection and nested PCR were used to identify tissue localizations of CFPHV in skin from a normal turtle. Of the turtles sampled in Manglar Bay, 30.5% had tumors; at the relatively more pristine Culebrita, 5.3% of turtles sampled had tumors. All three PCR primer combinations successfully amplified CFPHV from tumors, and from normal skin of both tumored and tumor-free turtles. Via nested PCR, the polymerase gene target proved superior to the other two gene targets in the positive detection of CFPHV DNA. CFPHV infection may be common relative to disease incidence, supporting the idea that extrinsic and/or host factors could play a transforming role in tumor expression. Laser capture microdissection revealed CFPHV in skin from a tumor-free turtle, harbored in both epidermal and dermal tissues. Identification of CFPHV harbored in a non-epidermal site (dermis) of a tumor-free turtle indicates that virus is latent in a non-tumored host.

Primary lacrimal gland adenocarcinoma of the third eyelid in a horse

A 5-year-old Draft Horse gelding presented for evaluation of a large, fleshy, ulcerated third eyelid mass OD of 3 weeks duration. Complete ophthalmic examination, ocular ultrasound and skull radiographs revealed a large soft-tissue mass involving the entire third eyelid OD and extending into the ventral right orbit to the level of the globe equator. No other abnormalities were noted on physical or ophthalmic examination. Surgical removal via exenteration was performed 3 months after initial presentation. A lacrimal adenocarcinoma of the third eyelid was diagnosed based on histopathology. Concurrent asymptomatic intra-ductal and intra-acinar Demodex caballi parasites were found in the eyelid sebaceous glands, likely as an incidental finding. No tumor recurrence or metastasis has occurred 12 months after excision. To the authors knowledge, this case is the first reported primary lacrimal adenocarcinoma in a horse. Complete surgical excision was curative.

Effects of graded doses of propofol for anesthesia induction on cardiovascular parameters and intraocular pressures in normal dogs

OBJECTIVE To determine the effects of graded doses of propofol on cardiovascular parameters and intraocular pressures (IOP) in normal dogs. STUDY DESIGN Prospective, randomized, modified Latin square, cross-over experimental study. ANIMALS Eleven adult random-source dogs weighing 20.2 +/- 5.7 kg. METHODS There were three treatment groups: propofol 8 mg kg(-1) intravenous (i.v.) until loss of jaw tone (Group P), propofol until loss of jaw tone +20% (Group P20), and propofol until loss of jaw tone +50% (Group P50). Atracurium 0.1 mg kg(-1) i.v. was administered in all treatments immediately after the propofol. All dogs received the three treatments in a randomized order, with at least a one week interval between treatments. Direct arterial blood pressure and IOP by applanation tonometry were obtained at baseline, after 5 minutes of pre-oxygenation (before induction), before, and after intubation. Blood gas samples were obtained at baseline, after pre-oxygenation, and before intubation. RESULTS There was no significant difference in IOP readings at any time point among groups. The IOP was significantly higher before intubation versus before induction in all three groups. There was a significantly smaller change in systolic, mean (MAP), and diastolic (DAP) arterial pressures in the P50 group compared with the P group after intubation. There was a significantly smaller change in MAP and DAP in the P50 group compared with the P20 group after intubation. The increase in CO(2) from before induction to before intubation was significantly greater in the P50 group than in the P or P20 groups. CONCLUSIONS AND CLINICAL RELEVANCE Graded doses of propofol did not affect the increase in IOP observed with propofol induction in normal dogs. Higher doses of propofol are of no apparent additional benefit in animals who cannot tolerate an abrupt increase in IOP but may be of benefit in dogs who cannot tolerate an abrupt increase in blood pressure accompanying orotracheal intubation.

Expression of Toll-like receptors 2, 3, 4, 6, 9, and MD-2 in the normal equine cornea, limbus, and conjunctiva

OBJECTIVE Human corneal cells have detectable levels of TLRs 1-10. TLRs 2 and 4 are the major corneal receptors, recognizing the PAMPs associated with fungal invasion in humans. The conjunctiva and limbus contain TLRs 2, 4, and 9. Our purpose was to determine the expression of TLRs 2, 3, 4, 6, 9, and MD-2 in the normal equine cornea, conjunctiva, and limbus. METHODS Corneal, limbal, and conjunctival tissues were collected from seven euthanized horses having no evidence of ocular disease. RNA extraction with DNase-1 digestion was performed followed by RT-PCR to determine expression of TLRs 2, 3, 4, 6, 9, and MD-2. Products were resolved by electrophoresis on 1.5% agarose gels and visualized using ethidium bromide staining. RESULTS Expression of TLRs 2, 3, 4, 6, 9, and MD-2 was present in the cornea, limbus, and conjunctiva of each horse, except one horse, where TLR3 expression was unable to be demonstrated in the dorsal and ventral conjunctiva. CONCLUSIONS Confirming the expression of TLRs in equine ocular tissues is an initial step in identifying how they play a role in infectious keratitis, particularly fungal. The results further support the use of equine ocular tissues as a model for human fungal keratitis. Studies of the TLR expression together with their cytokine profile induced during equine fungal keratitis may help further clarify the pathogenesis of the disease and possibly lead to the development of new treatment protocols for both equines and humans.

Diagnosis and management of chronic corneal epithelial defects (indolent corneal ulcerations)

Chronic corneal epithelial defects (CCEDs; indolent corneal ulcerations) are the most common refractory ulcerations in veterinary medicine and are diagnosed by their classic appearance. CCEDs are superficial ulcerations without stromal involvement and have a nonadherent epithelial border (lip). Fluorescein stain adheres to the exposed stroma and extends below the epithelial border, outlining the epithelial lip. CCEDs occur secondary to adnexal disease, keratoconjunctivitis sicca, exposure keratitis, neurotrophic keratitis, and primary corneal disease. In cats, herpes keratitis is associated with the development of CCEDs. Bacterial infections are not responsible for the refractory nature of CCEDs. Because of the refractory nature of CCEDs, treatment can be frustrating for both owner and veterinarian. Current treatment recommendations consist of identifying and treating the underlying cause and performing procedures that stimulate epithelialization and adhesion of the corneal epithelium. Initial treatment of CCEDs includes ulcer debridement and grid keratotomy. Superficial keratectomy is indicated in refractory cases.

Localization of Fibropapilloma-associated Turtle Herpesvirus in Green Turtles (Chelonia mydas) by In-Situ Hybridization

Fibropapilloma-associated turtle herpesvirus (FPTHV) is the presumed aetiological agent of sea turtle fibropapillomatosis (FP). Intralesional DNA and RNA of the virus have been detected by polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR), respectively, but the exact location and distribution of the virus within the tumours have not been addressed. In this study, in-situ hybridization (ISH) was used to investigate viral transcriptional activity and localization of FPTHV. Twenty-five tumours were obtained from the skin or conjunctiva of 105 green turtles (Chelonia mydas) examined on two islands in Puerto Rico (Culebra and Culebrita). These lesions comprised 19 fibropapillomas and six fibromas. FPTHV mRNA transcripts were detected by ISH in three fibropapillomas, with positive reactions confined to the nuclei of clusters of epithelial cells. Viral DNA was detected by riboprobe ISH combined with denaturation in 14 tumours, including both fibropapillomas and fibromas. Signals were confined to the nuclei of acanthotic epithelial cells and were not seen in the subepithelial fibrous areas of the tumours. These results suggest that FPTHV is present in epithelial cells and transcriptionally active in fibropapillomas.

Immunohistochemical study of matrix metalloproteinases-2 and -9, macrophage inflammatory protein-2 and tissue inhibitors of matrix metalloproteinases-1 and -2 in normal, purulonecrotic and fungal infected equine corneas.

OBJECTIVE Determine the effects of matrix metalloproteinases (MMPs)-2, -9, macrophage inflammatory protein-2 (MIP-2), tissue inhibitors of matrix metalloproteinase (TIMP)-1 and -2 by immunohistochemical expression in fungal affected and purulonecrotic corneas. PROCEDURE Paraffin-embedded equine corneal samples; normal (n = 9), fungal affected (FA; n = 26), and purulonecrotic without fungi (PN; n = 41) were evaluated immunohistochemically for MMP-2, -9, MIP-2, TIMP-1 and -2. The number of immunoreactive inflammatory cells was counted and statistics analyzed. Western blot was performed to detect MMP-2, MMP-9, TIMP-1 and TIMP-2 proteins. RESULTS Matrix metalloproteinases-2, -9, MIP-2, TIMP-1 and -2 immunoreactivity was identified in corneal epithelium of normal corneas, and in corneal epithelium, inflammatory cells, keratocytes, and vascular endothelial cells of both FA and PN samples. Inflammatory cell immunoreactivity was significantly higher in FA and PN samples than in the normal corneas. There was positive correlation between MMP-2 and MIP-2, MMP-9 and MIP-2, and MMP-9 and TIMP-1 in inflammatory cell immunoreactivity in FA samples. There was positive correlation between MMP-9 and MIP-2, MMP-9 and TIMP-2, MIP-2 and TIMP-1, and MIP-2 and TIMP-2 in inflammatory cell immunoreactivity in PN samples. Western blot confirmed the presence of all four proteins in equine corneal samples. CONCLUSION Increased immunoreactivity of MMP-2 and -9 in FA and PN samples is indirectly related to MIP-2 through its role in neutrophil chemo-attraction. Tissue inhibitors of matrix metalloproteinase-1 and TIMP-2 are up-regulated in equine purulonecrotic and fungal keratitis secondary to MMP-2 and MMP-9 expression. The correlation between MMPs -2 and -9, MIP-2, TIMPs -1 and -2 suggests that these proteins play a specific role in the pathogenesis of equine fungal keratitis.

Outcome of conjunctival flap repair for corneal defects with and without an acellular submucosa implant in 73 canine eyes.

OBJECTIVE To report and compare the success rate of a conjunctival pedicle flap (CPF) alone vs. a CPF with an underlying acellular submucosa implant for the repair of deep or perforating corneal wounds in dogs. PROCEDURES Records of 69 dogs (73 eyes) receiving a CPF with or without an acellular submucosa implant between 2004 and 2012 were reviewed. Successful outcome was defined as a comfortable eye with vision at the last post-operative evaluation. Age, breed, underlying corneal disease, surgical time, lesion characteristics, topical therapies, and postoperative complications were investigated. RESULTS Groups consisted of dogs that had a CPF alone (n = 37) and dogs that had a CPF plus an acellular submucosa implant (n = 36). Age, lesion size, surgical time, and time to discontinuation of topical anti-proteolytic medications was not significant between groups. Topical antibiotic use was terminated 13 days sooner (P ≤ 0.01) in dogs with an acellular submucosa implant. The combined success rate of all corneal wounds was 93% with success rate of corneal perforations, descemetoceles, and deep stromal wounds being 89%, 95%, and 100%, respectively. There was no difference in overall success rate between groups. Increasing age was associated with a negative outcome (P ≤ 0.01). Lesion size, presence of a corneal perforation, and concurrent keratoconjunctivitis sicca was not associated with a negative outcome. CONCLUSIONS A comparable success rate is achieved for deep or perforating corneal wounds stabilized with a CPF alone vs. a CPF plus acellular submucosa. Glaucoma, persistent uveitis, and cataract formation were not reported as post-operative complications in this study population.